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Laboratory (NREL) at www.nrel.gov/publications.

Contract No. DE-AC36-08GO28308

Novel Biological Conversion of 

Hydrogen and Carbon Dioxide 

Directly into Biodiesel

Cooperative Research and 

Development Final Report

CRADA Number: CRD-10-408

NREL Technical Contact: Pin-Ching Maness

CRADA Report

NREL/TP-2700-62148

June 2014
NOTICE

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Cover Photos: (left to right) photo by Pat Corkery, NREL 16416, photo from SunEdison, NREL 17423, photo by Pat Corkery, NREL 

16560, photo by Dennis Schroeder, NREL 17613, photo by Dean Armstrong, NREL 17436, photo by Pat Corkery, NREL 17721.

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This report is available at no cost from the National Renewable Energy Laboratory (NREL) at www.nrel.gov/publications.

Cooperative Research and Development Final Report

In accordance with Requirements set forth in Article XI.A(3) of the CRADA document, this document is 

the final CRADA report, including a list of Subject Inventions, to be forwarded to the Office of Science 

and Technical Information as part of the commitment to the public to demonstrate results of federally 

funded research.

CRADA Number: CRD-10-408

CRADA Title: Novel Biological Conversion of Hydrogen and Carbon Dioxide Directly into Biodiesel

Parties to the Agreement: OPX Biotechnologies

Joint Work Statement Funding Table Showing DOE Commitment: 

Estimated Costs NREL Shared Resources

Year 1 $ 636,485.00

Year 2 $ 718,778.00

Year 3 $ 534,007.00

TOTALS $ 1,889,270.00

Abstract of CRADA Work: 

OPX Biotechnologies, Inc. (OPX), the National Renewable Energy Laboratory (NREL), and Johnson 

Matthey will develop and optimize a novel, engineered microorganism that directly produces biodiesel 

from renewable hydrogen (H2) and carbon dioxide (CO2). The proposed process will fix CO2 utilizing H2

to generate an infrastructure-compatible, energy-dense fuel at costs of less than $2.50 per gallon, with 

water being produced as the primary byproduct. NREL will perform metabolic engineering on the 

bacterium Cupriavidus necator (formerly Ralstonia eutropha) and a techno-economic analysis to guide 

future scale-up work. H2 and CO2 uptakes rates will be genetically increased, production of free fatty 

acids will be enhanced and their degradation pathway blocked in order to meet the ultimate program goals.

Summary of Research Results: 

To improve H2 uptake, NREL had over-expressed the transcriptional regulator hoxA in C. necator. The 

outcomes lead to a two-fold improvement in the soluble hydrogenase activity and five-fold improvement 

in the membrane-bound hydrogenase activity. Moreover, we improved the CO2-fixation activity by 15-

fold via using a stronger promoter and deleting a terminator for the Rubisco enzyme when the changes 

were expressed from a plasmid. The changes have also been integrated into the chromosome leading to 

six-fold improvement. Both hydrogenase and Rubisco improvements have been integrated into a single C. 

necator host. Thus far all the improvements were verified based on in vitro assays. OPX is testing the 

effects of improved upstream substrate uptake on downstream enhanced production of fatty-acid fuels in 

scale-up reactors.

Subject Inventions Listing: None

Report Date: March 18, 2014
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This report is available at no cost from the National Renewable Energy Laboratory (NREL) at www.nrel.gov/publications.

Responsible Technical Contact at Alliance/NREL: Pin-Ching Maness

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